Methods of administering epothilone analogs for the treatment of cancer

ABSTRACT

A process for formulating certain epothilone analogs for parenteral administration is disclosed wherein the analog is dissolved in a mixture of at least 50% by volume tertiary-butanol in water, the mixture is lyophilized, the resulting lyophilized product is packaged in one vial with a sufficient amount of solvent comprising anhydrous ethanol and a suitable nonionic surfactant in a second vial. All steps are carried out with protection from light. In use, the contents of the second or diluent vial are added to the lyophilized product and mixed to constitute the epothilone analog and the resulting solution is diluted with a suitable diluent to produce a solution for intravenous injection containing the epothilone analog in a concentration of from about 0.1 mg/mL to about 0.9 mg/mL. A preferred surfactant is polyethoxylated castor oil and a preferred diluent is Lactated Ringer&#39;s Injection.

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority from provisional applicationserial Nos. 60/264,228, filed Jan. 25, 2001 and 60/290,006, filed May11, 2001, incorporated herein by reference in its entirety.

FIELD OF THE INVENTION

[0002] The present invention relates to methods of administration forparenteral and oral compositions of certain epothilone analogs that arecharacterized by enhanced clinical efficacy.

BACKGROUND OF THE INVENTION

[0003] Epothilones are macrolide compounds having utility in thepharmaceutical field. For example, Epothilones A and B having thestructures:

[0004] may be found to exert microtubule-stabilizing effects similar topaclitaxel (TAXOL®) and hence cytotoxic activity against rapidlyproliferating cells, such as, tumor cells or other hyperproliferativecellular disease, see Hofle et al., Angew. Chem. Int. Ed. Engl., Vol.35, No.13/14, 1567-1569 (1996); WO 93/10121 published May 27, 1993; andWO 97/19086 published May 29, 1997.

[0005] Derivatives and analogs of Epothilones A and B have beensynthesized and may be used to treat a variety of cancers and otherabnormal proliferative diseases. Such analogs are disclosed in Hofle etal., Id.; Nicolaou et al., Angew Chem. Int. Ed. Engl., Vol. 36, No. 19,2097-2103 (1997); and Su et al., Angew Chem. Int. Ed. Engl., Vol. 36,No. 19, 2093-2097 (1997).

[0006] Analogs of the epothilones that have been found to haveadvantageous activity are represented by formula I:

[0007] wherein the various symbols are as defined below. While thesecompounds possess significant therapeutic properties, they also presentdifficulties to those skilled in the art of pharmaceutical compounding,as a result of certain properties, as will be detailed hereinbelow. Inaccordance with the present invention, a formulation has been foundwhereby the epothilone analogs described above can be safely dispensedand administered via injection, without appreciable loss of potency.

[0008] Furthermore, many anti-cancer drugs have toxicity concerns.Indeed, the therapeutic profile of many potent antitumor drugs is pooras a result of toxicity. Therefore, there is also a need for methods ofadministration and dosing schedules that reduce or avoid the toxicityassociated with antitumor agents. The methods can allow exploitation ofpotent antitumor agents that would otherwise not be used clinically.

SUMMARY OF THE INVENTION

[0009] The invention encompasses a novel dosing schedule for epothilonecompounds, which schedule is useful in treating patients having solidtumors, particularly advanced solid tumors. Further, the methods of theinvention can be used to treat and/or prevent metastatic as well asprimary tumors. In one embodiment, the invention encompasses thetreatment of patients that have previously received either or bothradiation therapy and chemotherapy for solid tumors. It has also beenfound that the epothilone compounds of the invention particularly thepreferred compound,[1S-[1R*,3R*(E),7R*,10S*,11R*,12R*,16S*]]-7,11-Dihydroxy-8,8,10,12,16-pentamethyl-3-[1-methyl-2-(2-methyl-4-thiazolyl)ethenyl]-4-aza-17-oxabicyclo[14.1.0]heptadecane-5,9-dione,can be used to treat tumors refactory to radiation therapy orchemotherapy. The methods of the invention are useful against cancercells, and thus, tumors, that are naturally or become insensitive topaclitaxel.

[0010] In one embodiment, the dosing schedule of the invention comprisesthe weekly administration of an epothilone compound of the inventionpreferably as a one (1) hour infusion weekly on a continuous basis. Inanother embodiment, the administration is made weekly for a three weekcycle. The dose range for weekly infusion is from 1 mg/m² to 30 mg/m²and more preferably 1 mg/m² to 25 mg/m². In another embodiment, thedosing schedule includes both oral and intravenous administration of thesame epothilone compound. For example, the weekly infusion can befollowed or preceded by an oral administration of 20 mg/m² or greater.In a specific embodiment, the administration regimen includes a three(3) week cycle of intravenous infusion once per week for about one (1)hour followed by or preceded by an oral dose administered one or moretimes in the week before the first intravenous administration of a cycleor the week after the last intravenous administration of a cycle. Otherprotocols are also encompassed within the present invention includingbut not limited to:

[0011] (a) a daily dosing for 5 to 10 days followed by at least 3 daysof no dosing;

[0012] (b) weekly dosing for two to ten weeks followed by at least oneweek of no dosing; and

[0013] (c) dosing once every three weeks followed by at least one weekof no dosing.

[0014] The invention also contemplates the use of H₁ and H₂antihistamines before, after and/or before and after a cycle ofepothilone administration. Similarly, the invention encompasses the useof other chemotherapeutics, particularly anti-tumor agents, withepothilone cycle alone, or with the H₁ and H₂ blockers and theepothilones.

[0015] In another embodiment, the epothilone dosing schedule is usedafter the standard regimen of paclitaxel.

[0016] As discussed herein a wide variety of cancers are encompassed bythe methods of the present invention. In a preferred embodiment, themethods of the invention are for the treatment of solid tumors includingbut not limited to breast, head and neck, sarcoma, colorectal, UPT,melanoma, oesophagus, renal, cervix, thyroid, anal, ovarian, and colon.

[0017] The methods and compositions of the present invention describes aformulation and the preparation thereof for epothilone analogsrepresented by formula I:

[0018] wherein the various symbols are as defined below.

[0019] In one embodiment of the formulations of the present invention,the epothilone analog is initially solubilized with a mixture oftertiary-butanol and water and then lyophilized under optimizedconditions. The lyophilized drug is reconstituted first with a mixtureof a polyethoxylated castor oil surfactant and anhydrous ethanol, andthereafter diluted with Lactated Ringer's Injection to a concentrationappropriate for administration.

DETAILED DESCRIPTION OF THE INVENTION

[0020] In an embodiment, the present invention provides an advantageousformulation for the administration of epothilone analogs represented byformula I:

[0021] As used in formula I and throughout the specification, Q isselected from the group consisting of:

[0022] M is selected from the group consisting of oxygen, sulfur, NR⁸,and CR⁹R¹⁰;

[0023] each R¹, R², R³, R⁴, R⁵, R⁷, R¹¹, R¹², R¹³and R¹⁴ is,independently, selected from the group consisting of hydrogen, alkyl,substituted alkyl, aryl, substituted aryl and heterocyclo, and whereinR¹ and R² are alkyl, they can be joined to form cycloalkyl;

[0024] R⁶ is selected from the group consisting of hydrogen, alkyl,substituted alkyl, aryl, substituted aryl, cycloalkyl, heterocyclo andsubstituted heterocyclo;

[0025] R⁸ is selected from the group consisting of hydrogen, alkyl,substituted alkyl, R¹¹C═O, R¹²OC═O and R¹³SO₂; and

[0026] each R⁹ and R¹⁰ is, independently, selected from the groupconsisting of hydrogen, halogen, alkyl, substituted alkyl, aryl,heterocyclo, hydroxy, R¹⁴C═O, and R¹⁵OC═O.

[0027] The following are definitions of various terms used herein todescribe the present invention. These definitions apply to the terms asthey are used throughout this specification, unless otherwise limited inspecific instances, either individually or as part of a larger group.

[0028] The term “alkyl” refers to optionally substituted straight- orbranched-chain saturated hydrocarbon groups having from 1 to about 20carbon atoms, preferably from 1 to about 7 carbon atoms. The expression“lower alkyl” refers to optionally substituted alkyl groups having from1 to about 4 carbon atoms.

[0029] The term “substituted alkyl” refers to an alkyl group substitutedby, for example, one to four substituents, such as, halo,trifluoromethyl, trifluoromethoxy, hydroxy, alkoxy, cycloalkyoxy,heterocylooxy, oxo, alkanoyl, aryl, aryloxy, aralkyl, alkanoyloxy,amino, alkylamino, arylamino, aralkylamino, cycloalkylamino,heterocycloamino, disubstituted amino in which the two substituents onthe amino group are selected from alkyl, aryl, aralkyl, alkanoylamino,aroylamino, aralkanoylamino, substituted alkanoylamino, substitutedarylamino, substituted aralkanoylamino, thiol, alkylthio, arylthio,aralkylthio, cycloalkylthio, heterocyclothio, alkylthiono, arylthiono,aralkylthiono, alkylsulfonyl, arylsulfonyl, aralkylsulfonyl, sulfonamido(e.g., SO₂NH₂), substituted sulfonamido, nitro, cyano, carboxy, carbamyl(e.g., CONH₂), substituted carbamyl (e.g., CONH alkyl, CONH aryl, CONHaralkyl or instances where there are two substituents on the nitrogenselected from alkyl, aryl or aralkyl), alkoxycarbonyl, aryl, substitutedaryl, guanidino and heterocyclos, such as, indolyl, imidazolyl, furyl,thienyl, thiazolyl, pyrrolidyl, pyridyl, pyrimidyl and the like.Wherein, as noted above, the substituents themselves are furthersubstituted, such further substituents are selected from the groupconsisting of halogen, alkyl, alkoxy, aryl and aralkyl. The definitionsgiven herein for alkyl and substituted alkyl apply as well to the alkylportion of alkoxy groups.

[0030] The term “halogen” or “halo” refers to fluorine, chlorine,bromine and iodine.

[0031] The term “ring system” refers to an optionally substituted ringsystem containing one to three rings and at least one carbon to carbondouble bond in at least one ring. Exemplary ring systems include, butare not limited to, an aryl or a partially or fully unsaturatedheterocyclic ring system, which may be optionally substituted.

[0032] The term “aryl” refers to monocyclic or bicyclic aromatichydrocarbon groups having from about 6 to about 12 carbon atoms in thering portion, for example, phenyl, naphthyl, biphenyl and diphenylgroups, each of which may be substituted.

[0033] The term “aralkyl” refers to an aryl group bonded to a largerentity through an alkyl group, for example, a benzyl group.

[0034] The term “substituted aryl” refers to an aryl group substitutedby, for example, one to four substituents such as alkyl; substitutedalkyl, halo, trifluoromethyl, trifluoromethoxy, hydroxy, alkoxy,cycloalkyloxy, heterocyclooxy, alkanoyl, alkanoyloxy, amino, alkylamino,dialkylamino, aralkylamino, cycloalkylamino, heterocycloamino,alkanoylamino, thiol, alkylthio, cycloalkylthio, heterocyclothio,ureido, nitro, cyano, carboxy, carboxyalkyl, carbamyl, alkoxycarbonyl,alkylthiono, arylthiono, alkysulfonyl, sulfonamido, aryloxy and thelike. The substituent may be further substituted by one or more membersselected from the group consisting of halo, hydroxy, alkyl, alkoxy,aryl, substituted alkyl, substituted aryl and aralkyl.

[0035] The term “cycloalkyl” refers to optionally substituted saturatedcyclic hydrocarbon ring systems, preferably containing 1 to 3 rings and3 to 7 carbons per ring, which may be further fused with an unsaturatedC₃-C₇ carbocyclic ring. Exemplary groups include cyclopropyl,cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl,cyclodecyl, cyclododecyl, and adamantyl. Exemplary substituents includeone or more alkyl groups as described above, or one or more of thegroups described above as substituents for alkyl groups.

[0036] The terms “heterocycle”, “heterocyclic” and “heterocyclo” referto an optionally substituted, unsaturated, partially saturated, or fullysaturated, aromatic or nonaromatic cyclic group, for example, which is a4 to 7 membered monocyclic, 7 to 11 membered bicyclic, or 10 to 15membered tricyclic ring system, which has at least one heteroatom in atleast one carbon atom-containing ring. Each ring of the heterocyclicgroup containing a heteroatom may have 1, 2 or 3 heteroatoms selectedfrom nitrogen atoms, oxygen atoms and sulfur atoms, where the nitrogenand sulfur heteroatoms may also optionally be oxidized and the nitrogenheteroatoms may also optionally be quaternized. The heterocyclic groupmay be attached at any heteroatom or carbon atom.

[0037] Exemplary monocyclic heterocyclic groups include pyrrolidinyl,pyrrolyl, indolyl, pyrazolyl, oxetanyl, pyrazolinyl, imidazolyl,imidazolinyl, imidazolidinyl, oxazolyl, oxazolidinyl, isoxazolinyl,isoxazolyl, thiazolyl, thiadiazolyl, thiazolidinyl, isothiazolyl,isothiazolidinyl, furyl, tetrahydrofuryl, thienyl, oxadiazolyl,piperidinyl, piperazinyl, 2-oxopiperazinyl, 2-oxopiperidinyl,2-oxopyrrolidinyl, 2-oxazepinyl, azepinyl, 4-piperidonyl, pyridyl,N-oxo-pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, tetrahydropyranyl,tetrahydrothiopyranyl, tetrahydrothiopyranyl sulfone, morpholinyl,thiomorpholinyl, thiomorpholinyl sulfoxide, thiomorpholinyl sulfone,1,3-dioxolane and tetrahydro-1, 1-dioxothienyl, dioxanyl,isothiazolidinyl, thietanyl, thiiranyl, triazinyl, and triazolyl, andthe like.

[0038] Exemplary bicyclic heterocyclic groups include benzothiazolyl,benzoxazolyl, benzothienyl, quinuclidinyl, quinolinyl,quinolinyl-N-oxide, tetrahydroisoquinolinyl, isoquinolinyl,benzimidazolyl, benzopyranyl, indolizinyl, benzofuryl, chromonyl,coumarinyl, cinnolinyl, quinoxalinyl, indazolyl, pyrrolopyridyl,furopyridinyl (such as furo[2,3-c]pyridinyl, furo[3,1-b]pyridinyl] orfuro[2,3-b]pyridinyl), dihydroisoindolyl, dihydroquinazolinyl (such as3,4-dihydro-4-oxo-quinazolinyl), benzisothiazolyl, benzisoxazolyl,benzodiazinyl, benzofurazanyl, benzothiopyranyl, benzotriazolyl,benzpyrazolyl, dihydrobenzofuryl, dihydrobenzothienyl,dihydrobenzothiopyranyl, dihydrobenzothiopyranyl sulfone,dihydrobenzopyranyl, indolinyl, isochromanyl, isoindolinyl,naphthyridinyl, phthalazinyl, piperonyl, purinyl, pyridopyridyl,quinazolinyl, tetrahydroquinolinyl, thienofuryl, thienopyridyl,thienothienyl, and the like.

[0039] Exemplary substituents for the terms “ring system,”“heterocycle,” “heterocyclic,” and “heterocyclo” include one or moresubstituent groups as described above for substituted alkyl orsubstituted aryl, and smaller heterocyclos, such as, epoxides,aziridines and the like.

[0040] The term “alkanoyl” refers to —C(O)-alkyl.

[0041] The term “substituted alkanoyl” refers to —C(O)-substitutedalkyl.

[0042] The term “heteroatoms” shall include oxygen, sulfur and nitrogen.

[0043] The compounds represented by formula I form salts with a varietyof organic and inorganic acids. Such salts include those formed withhydrogen chloride, hydrogen bromide, methanesulfonic acid,hydroxyethanesulfonic acid, sulfuric acid, acetic acid, trifluoroaceticacid, maleic acid, benzenesulfonic acid, toluenesulfonic acid andvarious others as are recognized by those of ordinary skill in the artof pharmaceutical compounding. Such salts are formed by reacting acompound represented by formula I in an equivalent amount of the acid ina medium in which the salt precipitates or in an aqueous medium followedby evaporation.

[0044] In addition, zwitterions (“inner salts”) can be formed and areincluded within the term salts as used herein.

[0045] A particularly preferred epothilone analog within thoserepresented by formula I is [1S-[1R*,3R*(E),7R*,10S*,11R*,12R*,16S*,]]-7,11-Dihydroxy-8,8,10,12,16-pentamethyl-3-[1-methyl-2-(2-methyl-4-thiazolyl)ethenyl]-4-aza-7-oxabicyclo[14.1.0]heptadecane-5,9-dionerepresented by formula II:

[0046] The compounds represented by formulae I and II above, alsoreferred to herein as “the epothilone compounds of the invention,” andtheir preparation are described in U.S. patent application Ser. No.09/170,582, filed Oct. 13, 1998, and U.S. patent application Ser. No.09/280,191, filed Mar. 29, 1999, the disclosure of which is incorporatedherein by reference. The compounds represented by formulae I and IIabove may exist as multiple optical, geometric, and stereoisomers. Whilethe compounds shown herein are depicted for one optical orientation,included within the present invention are all isomers and mixturesthereof.

[0047] The compounds represented by formulae I and II above aremicrotubule-stabilizing agents. They are thus useful in the treatment ofa variety of cancers and other proliferative diseases including, but notlimited to, the following:

[0048] carcinoma, including that of the bladder, breast, colon, kidney,liver, lung, ovary, pancreas, stomach, cervix, thyroid and skin,including squamous cell carcinoma;

[0049] hematopoietic tumors of lymphoid lineage, including leukemia,acute lymphocytic leukemia, acute lymphoblastic leukemia, B-celllymphoma, T-cell lymphoma, Hodgkins lymphoma, non-Hodgkins lymphoma,hairy cell lymphoma and Burketts lymphoma;

[0050] hematopoietic tumors of myeloid lineage, including acute andchronic myelogenous leukemias and promyelocytic leukemia;

[0051] tumors of mesenchymal origin, including fibrosarcoma andrhabdomyoscarcoma;

[0052] other tumors, including melanoma, seminoma, teratocarcinoma,neuroblastoma and glioma;

[0053] tumors of the central and peripheral nervous system, includingastrocytoma, neuroblastoma, glioma, and schwannomas;

[0054] tumors of mesenchymal origin, including fibrosarcoma,rhabdomyoscaroma, and osteosarcoma; and

[0055] other tumors, including melanoma, xeroderma pigmentosum,keratoacanthoma, seminoma, thyroid follicular cancer andteratocarcinoma.

[0056] The compounds represented by formulae I and II are useful fortreating patients who have been previously treated for cancer, as wellas those who have not previously been treated for cancer. Indeed, themethods and compositions of this invention can be used in first-line andsecond-line cancer treatments. Furthermore, the compounds represented byformulae I and II are useful for treating refractory cancers.

[0057] The compounds represented by formulae I and II above will alsoinhibit angiogenesis, thereby affecting the growth of tumors andproviding treatment of tumors and tumor-related disorders. Suchanti-angiogenesis properties of the compounds represented by formulae Iand II will also be useful in the treatment of other conditionsresponsive to anti-angiogenesis agents including, but not limited to,certain forms of blindness related to retinal vascularization,arthritis, especially inflammatory arthritis, multiple sclerosis,restinosis and psoriasis.

[0058] Compounds represented by formulae I and II will induce or inhibitapoptosis, a physiological cell death process critical for normaldevelopment and homeostasis. Alterations of apoptotic pathwayscontribute to the pathogenesis of a variety of human diseases. Compoundsrepresented by formulae I and II, as modulators of apoptosis, will beuseful in the treatment of a variety of human diseases with aberrationsin apoptosis including, but not limited to, cancer and precancerouslesions, immune response related diseases, viral infections,degenerative diseases of the musculoskeletal system and kidney disease.

[0059] Each of the compounds represented by formulae I and II may alsobe formulated or co-administered with other therapeutic agents that areselected for their particular usefulness in administering therapiesassociates with the aforementioned conditions. For example, each of thecompounds of formulae I and II may be formulated with agents to preventnausea, hypersensitivity, and gastric irritation, such as anti-emetics,and H₁ and H₂ antihistamines. The above therapeutic agents, whenemployed in combination with the compound of formulae I or II, may beused in those amounts indicated in the Physicians' Desk Reference (PDR)or as otherwise determined by one of ordinary skill in the art.

[0060] Furthermore, compounds of formulae I or II may be administered incombination with other anti-cancer and cytotoxic agents and treatmentsuseful in the treatment of cancer or other proliferative diseases.Especially useful are anti-cancer and cytotoxic drug combinationswherein the second drug chosen acts in a different manner or differentphase of the cell cycle, e.g., S phase, than the present compounds offormula I and II which exert their effects at the G₂-M phase. Exampleclasses of anti-cancer and cytotoxic agents include, but are not limitedto, alkylating agents, such as nitorgen mustards, alkyl sulfonates,nitrosoureas, ethylenimines, and triazenes; antimetabolites, such asfolate antagonists, purine analogues, and pyrimidine analogues;antibiotics, such as anthracyclines, bleomycins, mitomycin,dactinomycin, and plicamycin; enzymes, such as L-asparaginase;farnesyl-protein transferase inhibitors; hormonal agents, such asglucocorticoids, estrogens/antiestrogens, androgens/antiandrogens,progestins, and luteinizing hormone-releasing hormone anatagonists,octreotide acetate; microtubule-disruptor agents, such as ecteinascidinsor their analogs and derivatives; microtubule-stabilizing agents such aspaclitaxel (Taxol®), docetaxel (Taxotere®); plant-derived products, suchas vinca alkaloids, epipodophyllotoxins, taxanes; and topoisomeraseinhibitors; prenyl-protein transferase inhibitors; and miscellaneousagents such as, hydroxyurea, procarbazine, mitotane, hexamethylmelamine,platinum coordination complexes such as cisplatin and carboplatin; andother agents used as anti-cancer and cytotoxic agents such as biologicalresponse modifiers, growth factors; immune modulators, and monoclonalantibodies. Compounds represented by formulae I and II may also be usedin conjunction with radiation therapy.

[0061] Representative examples of these classes of anti-cancer andcytotoxic agents include, but are not limited to, mechlorethaminehydrochlordie, cyclophosphamide, chlorambucil, melphalan, ifosfamide,busulfan, carmustin, lomustine, semustine, streptozocin, thiotepa,dacarbazine, methotrexate, thioguanine, mercaptopurine, fludarabine,pentastatin, cladribin, cytarabine, fluorouracil, doxorubicinhydrochloride, daunorubicin, idarubicin, bleomycin sulfate, mitomycin C,actinomycin D, safracins, saframycins, quinocarcins, discodermolides,vincristine, vinblastine, vinorelbine tartrate, etoposide, teniposide,paclitaxel, tamoxifen, estramustine, estramustine phosphate sodium,flutamide, buserelin, leuprolide, pteridines, diyneses, levamisole,aflacon, interferon, interleukins, aldesleukin, filgrastim,sargramostim, rituximab, BCG, tretinoin, irinotecan hydrochloride,betamethosone, gemcitabine hydrochloride, altretamine, and topoteca andany analogs or derivatives thereof.

[0062] Preferred members of these classes include, but are not limitedto, paclitaxel, cisplatin, carboplatin, doxorubicin, carminomycin,daunorubicin, aminopterin, methotrexate, methopterin, mitomycin C,ecteinascidin 743, porfiromycin, 5-fluorouracil, 6-mercaptopurine,gemcitabine, cytosine arabinoside, podophyllotoxin or podophyllotoxinderivatives such as etoposide, etoposide phosphate or teniposide,melphalan, vinblastine, vincristine, leurosidine, vindesine, andleurosine.

[0063] Examples of anti-cancer and other cytotoxic agents include thefollowing: cyclin dependent kinase inhibitors as found in WO 99/24416;and prenyl-protein transferase inhibitors as found in WO 97/30992 and WO98/54966.

[0064] The compounds may also be administered with or after anti-cancerand cytotoxic agents that are neurotoxic, i.e., poisonous to the nervoussystem.

[0065] Without being bound by any theory regarding mechanism ormorphology, the compounds represented by formulae I and II may also beused to treat conditions other than cancer or other proliferativediseases. Such conditions include, but are not limited to viralinfections such as herpesvirus, poxvirus, Epstein-Barr virus, Sindbisvirus and adenovirus; autoimmune diseases such as systemic lupuserythematosus, immune mediated glomerulonephritis, rheumatoid arthritis,psoriasis, inflammatory bowel diseases and autoimmune diabetes mellitus;neurodegenerative disorders such as Alzeimer's disease, AIDS-relateddementia, Parkinson's disease, amyotrophic lateral sclerosis, retinitispigmentosa, spinal muscular atrophy and cerebellar degeneration; AIDS;myelodysplastic syndromes; aplastic anemia; ischemic injury associatedmyocardial infarctions; stroke and reperfusion injury; restenosis;arrhythmia; atherosclerosis; toxin-induced or alcohol induced liverdiseases; hematological diseases such as chronic anemia and aplasticanemia; degenerative diseases of the musculoskeletal system such asosteoporosis and arthritis; aspirin-sensitive rhinosinusitis; cysticfibrosis; multiple sclerosis; kidney diseases; and cancer pain.

[0066] The compounds represented by formulae I and II, particularly thelatter, are difficult to formulate in that they possess very lowsolubility in aqueous media, rapidly degrade in contact with aqueousmedia, are sensitive to low pH when in solution, are light sensitive,are “Class D” cytotoxic, and have exceptionally poor wettingcharacteristics. Any one or two of these characteristics might becompensated for in compounding a pharmaceutical formulation forintravenous administration, but the combination of all of them presentsa formidable challenge to the pharmaceutical compounding chemist. Giventhe constraint that materials to be utilized in compounding anintravenous formulation must be approved for intravenous administration,the formulation provided in accordance with the present inventionunexpectedly was found to be suitable for overcoming the properties ofthe subject epothilone analogs, as noted above, that make them difficultto formulate. Initially, because of the fact that the subject epothiloneanalogs are poorly soluble in aqueous media and, in fact, rapidlydegrade in contact therewith, it was decided that they should beformulated in lyophilized form.

[0067] It has been found that a suitable media to form a solution of thesubject compounds for lyophilization is a mixture of tertiary-butanoland water for injection. This mixture must be at least about 50% v/v,preferably from about 50% to about 80% v/v tertiary butanol to preventdegradation of the subject epothilone analogs. Further, due to theexceptionally poor wetting characteristics of the subject epothiloneanalogs, the initial solution must be effected utilizing a mixture of atleast about 60% v/v, preferably from about 60% to about 95 % v/v,tertiary butanol and water. Once the solution is made, the requisiteamount of water or tertiary-butanol-water mixture can be added toachieve the final concentration for lyophilization as stated above.

[0068] It has unexpectedly been found that the stability of the subjectepothilone analogs can be significantly enhanced by carrying out thepreparation of the solution at a temperature below ambient, preferablyfrom about 5 EC to about 15 EC, more preferably about 5 EC. Further,both the process of forming the solution and subsequent lyophilizationare to be carried out in vessels such that the epothilone analogs areprotected from exposure to light. It is also beneficial to carry out thelyophilization in comparatively small batches so that the epothiloneanalogs are exposed to an aqueous medium for a minimum amount of time.

[0069] The primary drying stage of lyophilization of the solution formedas described above is carried out at temperatures from about −10 EC toabout −40 EC, preferably about −25 EC, under high vacuum, ie., fromabout 50 millitorr to about 300 millitorr, preferably about 200millitorr, for an extended period, i.e., from about 24 hours to about 96hours, preferably about 48 hours. Lyophilization in this temperaturerange produces an amorphous product which is desirable for anintravenous preparation. Those of ordinary skill in the art willappreciate that conventional procedures, such as powder X-raydiffraction, can be utilized to confirm the amorphous nature of thelyophilized product.

[0070] The residual solvents in the product are removed by a secondarydrying stage that is carried out at comparatively low temperatures,i.e., from about 10 EC to about 30 EC, preferably about 25 EC, underhigh vacuum, i.e., from about 50 millitorr to about 300 millitorr,preferably about 150 millitorr for an extended period, i.e., from about24 hours to about 96 hours, preferably about 48 hours.

[0071] It has unexpectedly been found that the stability of lyophilizedepothilone analogs described herein are not enhanced by excipientscommonly utilized for such purposes, such as lactose, mannitol, dextranand the like. Certain of these excipients may actually have a negativeeffect on the stability of the lyophilized product (lyophile). Hence,the epothilone analogs formulated in accordance with the presentinvention are lyophilized neat, i.e., without any excipient.

[0072] The lyophilized epothilone analogs represented by formulae I andII are reconstituted with a mixture of equal parts by volume ofDehydrated Alcohol, USP and a nonionic surfactant, preferably apolyoxyethylated castor oil surfactant available from GAF Corporation,Mount Olive, N.J., under the trademark, Cremophor EL. The lyophilizedproduct and vehicle for reconstitution are packaged separately inappropriately light-protected vials. To minimize the amount ofsurfactant in the reconstituted solution, only a sufficient amount ofthe vehicle is provided to form a solution having a concentration ofabout 2 mg/mL to about 4 mg/mL of the epothilone analog. Oncedissolution of the drug is achieved, the resulting solution is furtherdiluted prior to injection with a suitable parenteral diluent. Suchdiluents are well known to those of ordinary skill in the art. Thesediluents are generally available in clinical facilities. It is, however,within the scope of the present invention to package the subjectepothilone analogs with a third vial containing sufficient parenteraldiluent to prepare the final concentration for administration. Apreferred diluent is Lactated Ringer's Injection. The finalconcentration for administration would preferably contain from about 0.1mg/mL to about 0.9 mg/mL of the epothilone analog.

[0073] The final dilution of the reconstituted epothilone analog in theformulation of the invention may be carried out with other preparationshaving similar utility, for example, 5% Dextrose Injection, LactatedRinger's and Dextrose Injection, Sterile Water for Injection, and thelike. However, because of its narrow pH range, pH 6.0 to 7.5, LactatedRinger's Injection is preferred. Per 100 mL, Lactated Ringer's Injectioncontains Sodium Chloride USP 0.6 g, Sodium Lactate 0.31 g, Potassiumchloride USP 0.03 g and Calcium Chloride-2H20 USP 0.02g. The osmolarityis 275 mOsmol/L, which is very close to isotonicity.

[0074] The constituted preparation according to the present invention,i.e., the solution of the epothilone analog in the alcohol-surfactantvehicle, can be stored for up to about 24 hours before being furtherdiluted for administration. It has been found that the incidence ofallergic reactions encountered due to the presence of the surfactant inthe formulation is minimized by keeping its concentration at the minimumnecessary to effect solution of the epothilone analog. Further, theincidence of such reactions is about the same as has been experiencedwith other parenterally administered pharmaceuticals containing it, suchas cyclosporine. This observed level of allergic reaction with thepresent formulation is significantly lower that has been experiencedwith certain other oncology agents, such as Paclitaxel.

[0075] The present invention is also directed to methods of treatingcancer and other hyperproliferative diseases in patients comprisingadministering to the patient a therapeutically effective amount of oneor more compounds represented by formulae I and II. The compounds offormula I and II may be administered intravenously or orally, preferablyboth orally and intravenously. Preferably, the compounds of formulae Iand II are administered with one or more additional agents to preventnausea, hypersennsitivity, or gastric irritation such as an anti-emeticor an H₁ or H₂ antihistamine.

[0076] The amount of a compound represented by formulae I and IIadministered by each IV infusion, or orally, or both may be determinedby one of ordinary skill in the art, and includes exemplary dosageamounts for a human of from about 0.01 mg/kg/day to about 200 mg/kg/day,which may be administered in a single dose or in the form of individualdivided doses, such as from 1 to about 4 times per day. Preferably, thecompounds are administered in a dosage of less than about 100 mg/kg/day,and more preferably less than about 25 mg/kg/day in a single dose or inabout 2 to about 4 divided doses. It will be understood that thespecific dose level and frequency of dosage for any particular subjectmay be varied and will depend upon a variety of factors including theactivity of the specific compound employed, the metabolic stability andlength of action of that compound, the species, age, body weight,general health, sex and diet of the subject, the mode and time ofadministration, rate of excretion, drug combination, and severity of theparticular condition. Preferred subjects for treatment include animals,most preferably mammalian species such as humans, and domestic animalssuch as dogs, cats and the like, subject to the aforementioneddisorders.

[0077] Typically the compounds of formulae I and II are administereduntil the patient shows a response, for example, a reduction in tumorsize, or until dose limiting toxicity is reached. One or ordinary skillin the art will readily know when a patient shows a response or whendose limiting toxicity is reached. The common dose limiting toxicitiesassociated with compounds of formulae I and II include, but are notlimited to, fatigue, arthralgia/myalgia, anorexia, hypersensitivity,neutropenia, thrombocytopenia, and neurotoxicity.

[0078] When administered intravenously, the compounds of formulae I andII are preferably administered using the formulations of the invention.Generally, the compounds of Formulae I and II are administered by IVinfusion over a period of from about 10 minutes to about 3 hours,preferably about 30 minutes to about 2 hours, more preferably about 45minutes to 90 minutes, and most preferably about 1 hour. Typically, thecompounds are administered intravenously in a dose of from about 0.5mg/m² to 65 mg/m², preferably about 1 mg/m² to 50 mg/m², more preferablyabout 2.5 mg/m² to 30 mg/m², and most preferably about 25 mg/m².

[0079] One of ordinary skill in the art would readily know how toconvert doses from mg/kg to mg/m2 given either or both the height and orweight of the patient (See, e.g.,http://www.fda.gov/cder/cancer/animalframe.htm).

[0080] When administered orally the compounds of formulae I and II arepreferably administered in combination with a pharmaceuticallyacceptable acid neutralizing buffer. The buffer neutralizes acid in thestomach of the patient so that the rate of decomposition of thecompounds of formulae I and II is sufficiently decreased so that theyremain in the gastrointestinal tract for sufficient time to be absorbed.The compounds of formulae I and II may also be administered with ananti-acid such as hydroxides of aluminum and magnesium; carbonates, suchas sodium carbonate and calcium carbonate; silicates; and phosphates toneutralize the acid in the stomach before during or after administrationof the compounds of formulae I and II.

[0081] As used herein, the term “pharmaceutically acceptable acidneutralizing buffer” refers to a combination of a pharmaceuticallyacceptable non-toxic acid and a pharmaceutically acceptable non-toxicsalt of an acid that when added to a solution provides a solution thatis more resistant to change of pH, compared to a solution without thebuffer, when acid or alkali is added to the solution. The term“pharmaceutically acceptable acid neutralizing buffer” also includescompounds, such as basic compounds, that when added to an acidicsolution neutralizes the acid and increases the pH of the solution.

[0082] In one embodiment of the invention, the compounds of formulae Iand II and the pharmaceutically acceptable acid neutralizing buffer areprovided in a single oral dosage form and are administeredsimultaneously. The single composition comprising the combination of thecompounds of formulae I and II may be administered as a solid oraldosage form (e.g., a tablet, capsule, or powder) or a liquid oral dosageform (e.g., a solution, suspension, or elixir). The solution orsuspension can be constituted just prior to administration using theappropriate solvents or cosolvents to dissolve the epothilone and thebuffer components.

[0083] For example, the compounds of formulae I and II and thepharmaceutically acceptable acid neutralizing buffer may be orallyadministered simultaneously as a solution of the epothilone of formula(I) or (II) dissolved in a liquid comprising propyleneglycol:ethanol:phosphate buffer (for example at 1M, about pH 8) in aratio of about 58:12:30, respectively.

[0084] The compounds of formulae I and II and the pharmaceuticallyacceptable acid neutralizing buffer can also be provided as separatedistinct pharmaceutical compositions and administered separately. Eachof which are administered as a solid oral dosage form or a liquid oraldosage form. When the compounds of formulae I and II and thepharmaceutically acceptable acid neutralizing buffer are administeredseparately, the pharmaceutically acceptable acid neutralizing buffer maybe orally administered before, after, or both before and after thecompounds of formulae I and II is administered. Preferably, thepharmaceutically acceptable acid neutralizing buffer is administeredboth before and after oral administration of the compounds of formulae Iand II, in an amount sufficient to neutralize the stomach acid. When thepharmaceutically acceptable acid neutralizing buffer is administeredbefore the compounds of formulae I and II it is administered withinabout 5 hours preferably within about 3 hours, more preferably withinabout 1 hour, and most preferably with about 10 minutes before thecompounds of formulae I and II is administered. When thepharmaceutically acceptable acid neutralizing buffer is administeredafter the compounds of formulae I and II it is administered within about5 hours, preferably within about 3 hours, more preferably within about 1hour, and most preferably within about 10 minutes after the compounds offormulae I and II is administered.

[0085] The compounds of formulae I and II can also be administered as anenteric coated pill or capsule to delay release of the epothilone untilafter the pharmaceutically effective acid neutralizing buffer isadministered. Enteric coated tablets and capsules are capsules coatedwith a substances that resist solution in a gastric fluid butdisintegrate in the intestine.

[0086] Typically, the pharmaceutically acceptable acid neutralizingbuffer is administered in an amount sufficient to deliver at least about20 milliequivalents of acid neutralization capacity, preferably at leastabout 30 milliequivalents of acid neutralization capacity, morepreferably at least about 40 milliequivalents of acid neutralizationcapacity, and most preferably at least about 50 milliequivalents of acidneutralization capacity. Typically, the pharmaceutically acceptable acidneutralizing buffer is administered as an aqueous solution having a pHof between about 5 to 9, preferably about 6 to 8.5, and more preferablyabout 7 to 8. Any pharmaceutically acceptable acid neutralizing bufferthat provides a solution having a pH in the desired range may be used inthe methods of the invention. Preferably, the pharmaceuticallyacceptable acid neutralizing buffer is a dibasic phosphate-monobasicphosphate buffer or a dibasic phosphate buffer-citric acid-citratebuffer.

[0087] For example, oral administration of the compounds of formulae Iand II can involve first orally administering to the patient thepharmaceutically acceptable acid neutralizing buffer as about 150 mL ofan aqueous solution comprising anhydrous dibasic sodium phosphate (about0.2 M), sodium citrate dihydrate (about 0.07 M), and anhydrous citricacid (about 0.008 M) at a pH of about 7.4; followed by oraladministration of the compounds of formulae I and II as a liquid dosageform in a propylene glycol:ethanol system having a ratio of about 80:20;followed by oral administration of another about 150 mL aqueous solutioncomprising anhydrous dibasic sodium phosphate (about 0.2 M), sodiumcitrate dihydrate (about 0.07 M), and anhydrous citric acid (about 0.008M) at a pH of about 7.4.

[0088] As discussed above, the compounds of formulae I and II can beadministered orally, intravenously, or both. In particular, the methodsof the invention encompass dosing protocols such as once a day for 2 to10 days, preferably every 3 to 9 days, more preferably every 4 to 8 daysand most preferably every 5 days. In one embodiment there is a period of3 days to 5 weeks, preferably 4 days to 4 weeks, more preferably 5 daysto 3 weeks, and most preferably 1 week to 2 weeks, in between cycleswhere there is no treatment. In another embodiment the compounds offormulae I or II can be administered orally, intravenously, or both,once a day for 3 days, with a period of preferably 1 week to 3 weeks inbetween cycles where there is no treatment. In yet another embodimentthe compounds of formulae I or II can be administered orally,intravenously, or both, once a day for 5 days, with a period ofpreferably 1 week to 3 weeks in between cycles where there is notreatment.

[0089] In one preferred embodiment the treatment cycle foradministration of the compounds of formulae I or II is once daily for 5consecutive days and the period between treatment cycles is from 2 to 10days, preferably one week.

[0090] The compounds of formulae I and II can also be administeredorally, intravenously, or both once every 1 to 10 weeks, preferablyevery 2 to 8 weeks, more preferably every 3 to 6 weeks, and even morepreferably every 3 weeks.

[0091] In another method of the invention, the compounds of formulae Iand II are administered in a 28 day cycle wherein the compound offormulae I and II are intravenously administered on days 1, 7, and 14and orally administered on day 21. Alternatively, the compounds offormulae I and II are administered in a 28 day cycle wherein thecompound of formulae I and II are orally administered on day 1 andintravenously administered on days 7, 14, and 28.

[0092] According to the methods of the invention, the compounds offormulae I and II are administered until the patient shows a response,for example, a reduction in tumor size, or until dose limiting toxicityis reached.

[0093] Many anti-cancer agents are neurotoxic, e.g., they are known tocause side effects of the central and peripheral nervous system. Thisinvention further encompasses the use of compounds of formulae I and IIin patients previously experiencing neurotoxicity with other anti-canceragents. Although, the compounds of the invention may also causeneurotoxicity at certain doses, the methods herein can be used to reduceor avoid such toxicity.

EXAMPLES

[0094] The following non-limiting example serves to illustrate thepractice of the present invention.

Example 1

[0095] IV Dosage Form

[0096][1S-[1R*,3R*(E),7R*,10S*,11R*,12R*,16S*]]-7,11-Dihydroxy-8,8,10,12,16-pentamethyl-3-[1-methyl-2-(2-methyl-4-thiazolyl)ethenyl]-4-aza-17-oxabicyclo[14.1.0]heptadecane-5,9-dione,9.86 g, was wetted/partially dissolved with 600 mL of a 9:1 mixture oftertiary butanol and Water for Injection USP which had been pre-cooledto 5 EC. Once the drug powder had become completely wetted, dissolutionwas completed by the addition of 600 mL of a 1:9 mixture of tertiarybutanol and Water for Injection and 766 mL of a 1:1 mixture of tertiarybutanol and Water for Injection which likewise had been pre-cooled to 5EC thereby making the final solution a 1:1 mixture. The dissolution wascarried out under protection from light.

[0097] The solution formed above was promptly lyophilized in a VirtisINOTOP lyophilizer at −16 EC under light protectant conditions over aperiod of 48 hours. The resultant lyophilized product (lyophile) wasthen further dried at 15 EC under high vacuum for 48 hours. Nodetectable degradation of the drug was observed during these procedures.The lyophile was packaged under sterile conditions into 30 mL vials,each containing 10 mg of drug and standard excess to allow forvial/needle/syringe loss.

[0098] The lyophile is reconstituted with 5.5 mL of a 1:1 volume mixtureof Dehydrated Alcohol USP and Cremophor EL®, which typically will besupplied with the drug in a separate vial, to achieve a final drugconcentration of 2 mg/mL. Once dissolution is effected by gentlyswirling the vial, the resultant solution is diluted to achieve aconcentration of 0.2 mg/mL by the addition of 9 mL of Lactated Ringer'sfor Injection for each milliliter of constituted drug product.

Example 2

[0099] IV Administration of Compound II

[0100] A total of 24 cancer patients (12 male and 12 female) receivedcompound II by IV administration to evaluate the maximum tolerated dose(MTD), the dose limiting toxicity (DLT), the pharmacokinitics andpharmacodynamics, and to evaluate the anti-tumor activity of compoundII. The median age (range) of the patients was 57 (34-74). 5 patientshad breast cancer, 5 patients had head and neck cancer, 2 patients hadsarcoma, 2 patients had colorectal cancer, 2 patients had UPT cancer, 2patients had melanoma, 2 patients had cancer of the esophagus, 1 patienthad renal cancer, 1 patient had cervical cancer, 1 patient had thyroidcancer, and 1 patient had anal cancer. 21 patients had received priorchemotherapy (18 patients received neurotoxic agents and 18 patientsreceived radiotherapy). The median number of prior chemotherapy linesincluding adjuvant (range) was 2 (1-3).

[0101] Patients were orally administered compound II on day 1 (for doselevels of 20 mg/m² and higher) followed by a 30 minute IV infusion ofcompound II every week starting on day 7. Patients were administeredcompound II at doses of 1, 2.5, 5, 10, 20, 25, and 30 mg/ m². Patientswere monitored during the course of treatment for dose limiting toxicity(DLT) The results of the study showed that compound II can beadministered weekly at doses up to 30 mg/ m² without severe toxicitybeing observed.

[0102] In a second study a total of 12 cancer patients (5 male and 7female) were orally administered compound II on day 1 followed by a 30minute IV infusion of compound II every week starting on day 7 at a doseof 25 mg/m² to evaluate neurotoxicity of compound II. The median age(range) of the patients was 51 (30-65). 4 patients had colorectalcancer, 3 patients had breast cancer, 2 patients had melanoma, 1 patienthad renal cancer, 1 patient had sarcoma, and 1 patient had ovary cancer.10 patients had received prior chemotherapy (6 patients receivedneurotoxic agents and 18 patients received radiotherapy). The mediannumber of prior chemotherapy lines including adjuvant (range) was 2(0-3). This study showed that compound II can be used to patients thathave had prior chemotherapy lines that include the use of neurotoxicanticancer agents. In patients that have had prior chemotherapy linesthat use neurotoxic anticancer agents, however, it is preferably, thatthe cumulative dose of compound II does not exceed about 200 mg/m² percycle.

[0103] The study further showed that breast tumors and colon tumors, inpatients previously treated with chemotherapy, responded to treatmentwith compound II. Specifically, breast cancer patients previouslytreated with adriamycin and taxotere with cyclophosphamide,5-fluorouracil, methotrexate adjuvant therapy; adriamycin and taxoterewith cyclophosphamide, 5-fluorouracil, methotrexate adjuvant therapy; oradriamycin, cyclophosphamide, 5-fluorouracil for metastatic cancerresponded to treatment with compound II. Patients with metastatic coloncancer previously treated with taxol and carboplatin; 5-fluorouracil andleucovorin; or irinotecan responded to treatment with compound II.

Example 3

[0104] Pharmacokinetics of Compound II Orally Administered to CancerPatients

[0105] Patients with Advanced Malignancies were administered Compound IIweekly as a 30-minute infusion (a course=3 intravenous weeklyadministrations). Patients received doses of 1, 2.5, 5, 10, 20, 25, or30 mg/m². Starting at the 20 mg/m² dose level, a single oral dose ofCompound II was given on day 6 in a vehicle of 80% propylene glycol and20% ethanol (v/v) followed by administration of a citrate/phosphatebuffer (22.5 gm) before course 1 to assess the absolute bioavailabilityof Compound II. The dose of oral Compound II administered on Day 6matched the dose of IV Compound II administered on day 1. Serial plasmasampling was obtained on day 6 and day 1 of course 1 to assesspharmacokinetics by an LC/MS/MS.

[0106] Samples were analyzed by adding an internal standard to 0.2 mL ofplasma sample, precipitating with acetone, and then extracting thesupernatant with 1-chlorobutane. The organic layer was removed andevaporated to dryness. The residue was reconstituted and injected intothe LC/MS/MS system. Chromatographic separation was achieved,isocratically, on a YMC ODS-AQ column (4.6×50 mm, 3:m) with a mobilephase of acetonitrile:0.01M ammonium acetate, pH 5.0 (65:35). Detectionwas by negative electrospray tandem mass spectrometry. The standardcurve, which ranged from 2 to 500 ng/mL for all analytes and was fittedto a 1/x weighted quadratic regression model.

[0107] Compound II for oral administration, 25 mg/vial, was supplied as“drug in bottle.” The vehicle (buffer) for constitution of Compound II,25 mg/vial, was a mixture of 80% propylene glycol and 20% ethanol (v/v).The propylene glycol/ethanol mixture was prepared by mixing 80 parts byvolume of propylene glycol and 20 parts by volume of ethanol in asuitable container and gently swirling the container until the solutionwas completely mixed.

[0108] The citrate/phosphate buffer for oral administration aftercompound II was supplied in a separate bottle. Buffer for use withCompound II was constituted with water for injection (WFI).

[0109] Compound II was prepared for administration to patients by usinga suitable syringe to slowly inject 2.5, 5, or 10 mL of the propyleneglycol/ethanol mixture into the 20 cc vial containing 25 mg/vial ofCompound II, to give concentrations of 10, 5, or 2.5 mg/mL,respectively, depending on the dose to be administered to the patient.The syringe was removed and the vial shaken vigorously for 10 seconds.The vial was placed in a sonication bath and sonicated until thesolution became clear. Vials were pooled depending on the dose.

[0110] The buffer for administration with Compound II was supplied in an8 oz. clear glass bottle and was constituted with water for injection(WFI). The child resistant cap was removed from the bottle of buffer andabout 140 mL of water for injection (WFI) were added. The bottle wasshaken vigorously or sonicated with intermittent shaking until a clearsolution was obtained.

[0111] Following oral administration on day 6, 7 mL blood samples wascollected into Becton Dickinson Vacutainer tubes with K3EDTA asanticoagulent (lavender-colored top) according to the following schedule(expressed as hours:minutes from the start of the oral administration):predose, 00:15, 00:30, 00:45, 1:00, 1:30, 2:00, 3:00, 4:00, 6:00, 8:00,24:00, 48:00, and 72:00. Following IV administration on day 1, 7 mLblood samples were collected into Becton Dickinson Vacutainer tubes withK3EDTA as anticoagulent (lavender-colored top) according to thefollowing schedule (expressed as hours:minutes from the start of the IVinfusion):predose, 00:15, 00:30 (end-of infusion), 00:45, 1:00, 1:30,2:00, 3:00, 4:00, 6:00, 8:00, 24:00, 48:00, and 72:00.

[0112] Immediately after blood collection, the Vacutainer tubes wereinverted several times to ensure mixing with the anticoagulant and thenimmediately placed on crushed ice. Within 30 minutes of collection,samples were centrifuged for 5 minutes at approximately 2000×g and 0 to5 EC. The plasma was then transferred to separate pre-labeledscrew-capped polypropylene tubes and stored at −70 EC until bioanalysis.Plasma concentrations of Compound II were analyzed using the LC/MS/MSassay.

[0113] The plasma concentration versus time data were analyzed usingnon-compartmental methods. The pharmacokinetic parameters determined forCompound II included the maximum observed plasma concentration (Cmax),time to reach Cmax (Tmax), area under the plasma concentration timecurve from time zero to the time of last sampling time T(AUC(0-T)).

[0114] A total of 18 patients have received oral Compound II as asolution on day 6 and by IV on day 1. The summary of the pharmacokineticresults from these patients is presented in Table 1. TABLE 1 Summary ofPharmacokinetics of Patients Administered Compound II Orally andIntravenously Dose (mg/m²) 20 25 30 N 3 11 4 Route IV Oral IV Oral IVOral Formulation IV Solution IV Solution IV Solution for Oral for Oralfor Oral Admin. Admin. Admin. CMAX^(a) 251 142 447 180 711 274 (ng/mL)(108) (106) (189) (110) (530) (104) TMAX^(b) 0.25 1.0 0.50 0.50 0.500.50 (h) (0.25, (0.25, (0.25, (0.25, (0.25, (0.25, 0.25) 1.50) 0.50)3.00) 0.50) 0.75) AUC(0-T)^(a,c) 796 404 848 533 1155 708 (H.ng/mL)(587) (381) (284) (577) (292) (291) % F^(a) NA 43.5 NA 55.6 NA 62.2(16.1) (18.4) (25.1)

[0115] The embodiments of the invention described above are intended tobe merely exemplary, and those skilled in the art will recognize, orwill be able to ascertain using no more than routine experimentation,numerous equivalents of specific compounds, materials, and procedures.All such equivalents are considered to be within the scope of theinvention and are encompassed by the appended claims.

What is claimed is:
 1. A process for formulating, for parenteraladministration, an epothilone analog represented by formula I:

wherein: Q is selected from the group consisting of:

M is selected from the group consisting of oxygen, sulfur, NR⁸, andCR⁹R¹⁰; each R¹, R², R³, R⁴, R⁵, R⁷, R¹¹, R¹², R¹³, R¹⁴ and R¹⁵ is,independently, selected from the group consisting of hydrogen, alkyl,substituted alkyl, aryl, substituted aryl and heterocyclo, and whereinR¹ and R² are alkyl, they can be joined to form cycloalkyl; R⁶ isselected from the group consisting of hydrogen, alkyl, substitutedalkyl, aryl, substituted aryl, cycloalkyl, heterocyclo and substitutedheterocyclo; R⁸ is selected from the group consisting of hydrogen,alkyl, substituted alkyl, R¹¹C═O, R¹²OC═O and R¹³SO₂; and each R⁹ andR¹⁰ is, independently, selected from the group consisting of hydrogen,halogen, alkyl, substituted alkyl, aryl, heterocyclo, hydroxy, R¹⁴C═O,and R¹⁵OC═O; and any salts, solvates, or hydrates thereof, comprisingthe following steps carried out under protection from light: a)dissolving said epothilone analog in a mixture of at least about 50% byvolume tertiary-butanol in water to form a solution; b) performingprimary drying of said solution at a temperature of from about −10 EC toabout −40 EC under high vacuum of from about 50 millitorr to about 300millitorr for from about 24 hours to about 96 hours to form alyophilized product; c) performing secondary drying of the resultantlyophilized product at a temperature of from about 10 EC to about 30 ECunder high vacuum of from about 50 millitorr to about 300 millitorr forfrom 24 hours to about 96 hours; and d) packaging said lyophilizedproduct in a first vial in combination with a second vial containing asufficient quantity of an equal mixture by volume of a suitable nonionicsurfactant and anhydrous ethanol to effect solution thereof.
 2. Theprocess of claim 1 wherein said epothilone analog is represented byformula II:


3. The process of claim 1 wherein in step a) said analog is first wettedwith a mixture of at least about 60% tertiary-butanol in water, and thensufficient water, or a mixture of tertiary-butanol and water, is addedthereto so that the resulting solution contains from about 2 mg/mL toabout 30 mg/mL of said analog in a mixture of from about 50% to about80% by volume tertiary-butanol in water.
 4. The process of claim 2wherein in step a) said analog is first wetted with a mixture of atleast about 60% tertiary-butanol in water, and then sufficient water, ora mixture of tertiary-butanol and water, is added thereto so that theresulting solution contains from about 2 mg/mL to about 30 mg/mL of saidanalog in a mixture of from about 50% to about 80% by volumetertiary-butanol in water.
 5. The process of claim 3 wherein in step a)said analog is initially wetted with a mixture of from about 60% toabout 95% by volume tertiary-butanol in water.
 6. The process of claim 4wherein in step a) said analog is initially wetted with a mixture offrom about 60% to about 95% by volume tertiary-butanol in water.
 7. Theprocess of claim 1 wherein said primary drying in step b) is carried outat a temperature of about −25° C. and a pressure of about 200 millitorrfor about 48 hours.
 8. The process of claim 2 wherein said primarydrying in step b) is carried out at a temperature of about −25° C. and apressure of about 200 millitorr for about 48 hours.
 9. The process ofclaim 1 wherein said secondary drying in step c) is carried out at atemperature of about 25° C. and a pressure of about 150 millitorr forabout 48 hours.
 10. The process of claim 2 wherein said secondary dryingin step c) is carried out at a temperature of about 25° C. and apressure of about 150 millitorr for about 48 hours.
 11. The process ofclaim 1 wherein said surfactant is polyethoxylated castor oil.
 12. Theprocess of claim 2 wherein said surfactant is polyethoxylated castoroil.
 13. The process of claim 11 wherein said second vial contains anamount of said mixture sufficient to form a solution of from about 2mg/mL to about 4 mg/mL of said analog therein.
 14. The process of claim12 wherein said second vial contains an amount of said mixturesufficient to form a solution of from about 2 mg/mL to about 4 mg/mL ofsaid analog therein.
 15. A pharmaceutical preparation comprising, inseparate vials, a lyophilized epothilone analog and a quantity of asolvent therefor such that when the contents of said vials are combined,the resulting solution contains from about 2 mg/mL to about 4 mg/mL ofsaid analog, said solvent comprising a mixture of about equal parts byvolume of dehydrated ethanol and a suitable nonionic surfactant, saidanalog being represented by formula I:

wherein: Q is selected from the group consisting of

M is selected from the group consisting of oxygen, sulfur, NR⁸, andCR⁹R¹⁰; each R¹, R², R³, R⁴, R⁵, R⁷, R¹¹, R¹², R¹³, R¹⁴ and R¹⁵ is,independently, selected from the group consisting of hydrogen, alkyl,substituted alkyl, aryl, substituted aryl and heterocyclo, and whereinR¹ and R² are alkyl, they can be joined to form cycloalkyl; R⁶ isselected from the group consisting of hydrogen, alkyl, substitutedalkyl, aryl, substituted aryl, cycloalkyl, heterocyclo and substitutedheterocyclo; R⁸ is selected from the group consisting of hydrogen,alkyl, substituted alkyl, R¹¹C═O, R¹² 0C═O and R¹³SO₂; and each R⁹ andR¹⁰ is, independently, selected from the group consisting of hydrogen,halogen, alkyl, substituted alkyl, aryl, heterocyclo, hydroxy, R¹⁴C═O,and R¹⁵C═O; and any salts, solvates, or hydrates thereof.
 16. Thepharmaceutical preparation of claim 15 wherein said epothilone analog isrepresented by formula II:


17. The pharmacetuicl preparation of claim 15 wherein said nonionicsurfactant is polyethoxylated castor oil.
 18. A process for forming apharmaceutical composition for parenteral administration comprisingmixing the contents of the vials of the pharmaceutical preparation ofclaim 15 to effect solution of said lyophilized epothilone analog anddiluting the resultant solution with a quantity of a suitable parenteraldiluent such that the concentration of said analog therein will be fromabout 0.1 mg/mL to about 0.9 mg/mL.
 19. A process for forming aphamaceutical composition for parenteral administration comprisingmixing the contents of the vials of the pharmaceutical preparation ofclaim 16 to effect solution of said lyophilized epothilone analog anddiluting the resulting solution with a quantity of a suitable parenteraldiluent such that the concentration of said analog therein will be fromabout 0.1 mg/mL to about 0.9 mg/mL.
 20. A process for forming apharmaceutical composition for parenteral administration comprisingmixing the contents of the vials of the pharmaceutical preparation ofclaim 17 to effect solution of said lyophilized epothilone analog anddiluting the resultant solution with a quantity of a suitable parenteraldiluent such that the concentration of said analog therein will be fromabout 0.1 mg/mL to about 0.9 mg/mL.
 21. The process of claim 18 whereinsaid diluent is Lactated Ringer's Injection.
 22. The process of claim 19wherein said diluent is Lactated Ringer's Injection.
 23. The process ofclaim 20 wherein said diluent is Lactated Ringer's Injection.
 24. Amethod for treating a patient in need of treatment with an epothiloneanalog represented formula I:

wherein: Q is selected from the group consisting of

M is selected from the group consisting of oxygen, sulfur, NR⁸, andCR⁹R¹⁰; each R¹, R², R³, R⁴, R⁵, R⁷, R¹¹, R¹², R¹³, R¹⁴ and R¹⁵ is,independently, selected from the group consisting of hydrogen, alkyl,substituted alkyl, aryl, substituted aryl and heterocyclo, and whereinR¹ and R² are alkyl, they can be joined to form a cycloalkyl; R⁶ isselected from the group consisting of hydrogen, alkyl, substitutedalkyl, aryl, substituted aryl, cycloalkyl, heterocyclo and substitutedheterocyclo; R⁸ is selected from the group consisting of hydrogen,alkyl, substituted alkyl, R¹¹C═O, R¹²OC═O and R¹³SO₂; and each R⁹ andR¹⁰ is, independently, selected from the group consisting of hydrogen,halogen, alkyl, substituted alkyl, aryl, heterocyclo, hydroxy, R¹⁴C═O,and R¹⁵OC═O; and any salts, solvates, or hydrates thereof, comprisingadministering to said patient, by intravenous injection, an effectiveamount of a pharmaceutical composition of claim
 18. 25. A method fortreating a patient in need of treatment with an epothilone analogrepresented formula I:

wherein: Q is selected from the group consisting of

M is selected from the group consisting of oxygen, sulfur, NR⁸, andCR⁹R¹⁰; each R¹, R², R³, R⁴, R⁵, R⁷, R¹¹, R¹², R¹³, R¹⁴ and R¹⁵ is,independently, selected from the group consisting of hydrogen, alkyl,substituted alkyl, aryl, substituted aryl and heterocyclo, and whereinR¹ and R² are alkyl, they can be joined to form a cycloalkyl; R⁶ isselected from the group consisting of hydrogen, alkyl, substitutedalkyl, aryl, substituted aryl, cycloalkyl, heterocyclo and substitutedheterocyclo; R⁸ is selected from the group consisting of hydrogen,alkyl, substituted alkyl, R¹¹C═O, R¹²OC═O and R¹³SO₂; and each R⁹ andR¹⁰ is, independently, selected from the group consisting of hydrogen,halogen, alkyl, substituted alkyl, aryl, heterocyclo, hydroxy, R¹⁴C═O,and R¹⁵OC═O; and any salts, solvates, or hydrates thereof, comprisingadministering to said patient, by intravenous injection, an effectiveamount of a pharmaceutical composition of claim
 19. 26. A method fortreating a patient in need of treatment with an epothilone analogrepresented formula I:

wherein: Q is selected from the group consisting of

M is selected from the group consisting of oxygen, sulfur NR⁸, andCR⁹R¹⁰; each R¹, R², R³, R⁴, R⁵, R⁷, R¹¹, R¹², R¹³, R¹⁴ and R¹⁵ is,independently, selected from the group consisting of hydrogen, alkyl,substituted alkyl, aryl, substituted aryl and heterocyclo, and whereinR¹ and R² are alkyl, they can be joined to form a cycloalkyl; R⁶ isselected from the group consisting of hydrogen, alkyl, substitutedalkyl, aryl, substituted aryl, cycloalkyl, heterocyclo and substitutedheterocyclo; R⁸ is selected from the group consisting of hydrogen,alkyl, substituted alkyl, R¹¹C═O, R¹²OC═O and R¹³SO₂; and each R⁹ andR¹⁰ is, independently, selected from the group consisting of hydrogen,halogen, alkyl, substituted alkyl, aryl, heterocyclo, hydroxy, R¹⁴C═O,and R¹⁵OC═O; and any salts, solvates, or hydrates thereof, comprisingadministering to said patient, by intravenous injection, an effectiveamount of a pharmaceutical composition of claim
 20. 27. The method ofclaim 24 wherein said diluent is Lactated Ringer's Injection.
 28. Themethod of claim 25 wherein said diluent is Lactated Ringer's Injection.29. The method of claim 26 wherein said diluent is Lactated Ringer'sInjection.
 30. A method of treating cancer in a patient comprisingintravenously and orally administering to said patient a therapeuticallyeffective amount of a compound represented by formula I:

wherein: Q is selected from the group consisting of

M is selected from the group consisting of oxygen, sulfur, NR⁸, andCR⁹R¹⁰; each R¹, R², R³, R⁴, R⁵, R⁷, R¹¹, R¹², R¹³, R¹⁴ and R¹⁵ is,independently, selected from the group consisting of hydrogen, alkyl,substituted alkyl, aryl, substituted aryl and heterocyclo, and whereinR¹ and R² are alkyl, they can be joined to form a cycloalkyl; R⁶ isselected from the group consisting of hydrogen, alkyl, substitutedalkyl, aryl, substituted aryl, cycloalkyl, heterocyclo and substitutedheterocyclo; R⁸ is selected from the group consisting of hydrogen,alkyl, substituted alkyl, R¹¹C═O, R¹²OC═O and R¹³SO²; and each R⁹ andR¹⁰ is, independently, selected from the group consisting of hydrogen,halogen, alkyl, substituted alkyl, aryl, heterocyclo, hydroxy, R¹⁴C═O,and R¹⁵OC═O; and any salts, solvates, or hydrates thereof.
 31. Themethod of claim 30, wherein the compound of formula I is administeredorally in a dose of about 0.05 mg/kg to 200 mg/kg.
 32. The method ofclaim 31, wherein the compound of formula I is administeredintravenously at a dose of about 1 mg/m² to 65 mg/m².
 33. The method ofclaim 32, wherein the compound of formula I is administeredintravenously at a dose of about 25 mg/m².
 34. The method of claim 30,wherein the compound of formula I is administered either orally orintravenously on a weekly basis.
 35. The method of claim 34, wherein a 3week cycle of intravenous administration is used.
 36. The method ofclaim 35, wherein the compound of formula I is administered orallybefore the 3 week cycle.
 37. The method of claim 35, wherein thecompound of formula I is administered orally after the 3 week cycle. 38.The method of claim 31, wherein the compound of formula I isadministered as one or more 28 day cycles, wherein the compound offormula I is administered as an IV infusion on days 1, 7, and 14 andorally on day
 21. 39. The method of claim 31, wherein the IV infusion isadministered over a period of about 45 minutes to 90 minutes.
 40. Themethod of claim 39, wherein the IV infusion is administered over aperiod of about 1 hour.
 41. The method of claim 31, further comprisingadministering to said patient one or more additional therapeutic agentsto prevent nausea, vomiting, hypersensitivity, or gastric irritation.42. The method of claim 41, wherein the one or more additionaltherapeutic agents is an H₁ or H₂ antihistamine.
 43. The method of claim31, wherein the patient has not previously been treated for cancer. 44.The method of claim 31, wherein the patient has been previously treatedfor cancer.
 45. The method of claim 31, wherein the cancer is refractoryto radiation therapy.
 46. The method of claim 31, wherein the cancer isrefractory to anti-cancer chemotherapy.
 47. A pharmaceutical compositionsuitable for parenteral administration comprising a compound representedby formula I:

wherein: Q is selected from the group consisting of

M is selected from the group consisting of oxygen, sulfur, NR⁸, andCR⁹R¹⁰; each R¹, R², R³, R⁴, R⁵, R⁷, R¹¹, R¹², R¹³, R¹⁴ and R¹⁵ is,independently, selected from the group consisting of hydrogen, alkyl,substituted alkyl, aryl, substituted aryl and heterocyclo, and whereinR¹ and R² are alkyl, they can be joined to form a cycloalkyl; R⁶ isselected from the group consisting of hydrogen, alkyl, substitutedalkyl, aryl, substituted aryl, cycloalkyl, heterocyclo and substitutedheterocyclo; R⁸ is selected from the group consisting of hydrogen,alkyl, substituted alkyl, R¹¹C═O, R¹²OC═O and R¹³SO₂; and each R⁹ andR¹⁰ is, independently, selected from the group consisting of hydrogen,halogen, alkyl, substituted alkyl, aryl, heterocyclo, hydroxy, R¹⁴C═O,and R¹⁵OC═O; and any salts, solvates, or hydrates thereof; dehydratedalcohol; and a non-ionic surfactant.
 48. The composition of claim 47,wherein the surfactant is polyethoxylated castor oil.
 49. Thecomposition of claim 47, wherein the surfactant is Cremophor EL®. 50.The composition of claim 47, wherein the concentration of the compoundof formula I is from about 2 mg/mL to 4 mg/mL.
 51. The composition ofclaim 47, wherein the compound of formula I is


52. A method of treating cancer in a patient comprising intravenouslyadministering to said patient a therapeutically effective amount of thepharmaceutical formulation of claim 47 diluted in a parenteral diluent.53. The method of claim 52, wherein the parenteral diluent is 5%dextrose, lactated ringer's and dextrose injection, or sterile water forinjection.
 54. The method of claim 52, wherein the concentration of thecompound of formula I in the parenteral diluent is about 0.1 mg/mL to0.9 mg/mL.
 55. The method of claim 52, wherein the compound of formula Iis administered in a dose of about 1 mg/m² to 65 mg/m².
 56. The methodof claim 55, wherein the compound of formula I is administered at a doseof about 25 mg/m².
 57. The method of claim 52, wherein thepharmaceutical composition is administered as weekly as an IV infusion.58. The method of claim 52, wherein the IV infusion is administered overa period of about 45 minutes to 90 minutes.
 59. The method of claim 52,wherein the IV infusion is administered over a period of about 1 hour.60. The method of claim 52, further comprising administering to saidpatient one or more additional agents to prevent nausea, vomiting,hypersensitivity, or gastric irritation.
 61. The method of claim 60,wherein the one or more additional agents is an H₁ or H₂ antihistamine.62. The method of claim 52, wherein the patient has not previously beentreated for cancer.
 63. The method of claim 52, wherein the patient hasbeen previously treated for cancer.
 64. The method of claim 52, whereinthe cancer is refractory to radiation therapy.
 65. The method of claim52, wherein the cancer is refractory to anti-cancer chemotherapy.
 66. Amethod of treating cancer in a patient previously experiencingneurotoxicity comprising intravenously administering to said patient atherapeutically effective amount of the pharmaceutical formulation ofclaim 47 diluted in a parenteral diluent as a weekly infusion, whereinthe total dose of the compound of formula I is less than about 200mg/m².
 67. The method of claim 52, wherein the cancer is a solid tumor.68. The method of claim 30, wherein the cancer is a solid tumor.
 69. Amethod of treating cancer while reducing or avoiding neurotoxicity whichcomprises intravenously infusing a therapeutically effective amount ofcompound represented by formula I:

wherein: Q is selected from the group consisting of

M is selected from the group consisting of oxygen, sulfur, NR⁸, andCR⁹R¹⁰; each R¹, R², R³, R⁴, R⁵, R⁷, R¹¹, R¹², R¹³, R¹⁴ and R¹⁵ is,independently, selected from the group consisting of hydrogen, alkyl,substituted alkyl, aryl, substituted aryl and heterocyclo, and whereinR¹ and R² are alkyl, they can be joined to form a cycloalkyl; R⁶ isselected from the group consisting of hydrogen, alkyl, substitutedalkyl, aryl, substituted aryl, cycloalkyl, heterocyclo and substitutedheterocyclo; R⁸ is selected from the group consisting of hydrogen,alkyl, substituted alkyl, R¹¹C═O, R¹²OC═O and R¹³SO₂; and each R⁹ andR¹⁰ is, independently, selected from the group consisting of hydrogen,halogen, alkyl, substituted alkyl, aryl, heterocyclo, hydroxy, R¹⁴C═O,and R¹⁵OC═O; and any salts, solvates, or hydrates thereof; over a periodof one (1) hour to a patient in need thereof.
 70. The method of claim69, wherein the infusion is made on a weekly basis.
 71. The method ofclaim 69, wherein the therapeutically effective amount is from about 1mg/m² to about 65 mg/m².
 72. The method of claim 71, wherein the amountis 25 mg/m².
 73. The method of claim 69, wherein the compound of formulaI is


74. The method of claim 69 which further comprises orally administeringsaid compound 1 week before or after an intravenous administration. 75.A method of treating cancer in a human patient in need thereof with asynthetic or semi-synthetic epothilone analogue that is active againstcancer which comprises a four (4) week dosing cycle wherein said cyclecomprises three weeks of weekly intravenous administration and one weekof oral administration of said epothilone analogue.
 76. The method ofclaim 31, wherein the compound is administered daily or weekly.
 77. Themethod of claim 32, wherein the compound is administered daily orweekly.
 78. The method of claim 76 wherein the compound is administereddaily for 3 days with a period of 1 week to 3 weeks between cycles wherethere is no administration of the compound.
 79. The method of claim 77wherein the compound is administered daily for 3 days with a period of 1week to 3 weeks between cycles where there is no administration of thecompound.
 80. The method of claim 76 wherein the compound isadministered daily for 5 days with a period of 1 week to 3 weeks betweencycles where there is no administration of the compound.
 81. The methodof claim 77 wherein the compound is administered daily for 5 days with aperiod of 1 week to 3 weeks between cycles where there is noadministration of the compound.
 82. The method of claim 31 wherein thecompound of formula I is administered as one or more 28 day cycles,wherein the compound of formula I is administered orally on day 1 and asan IV infusion on days 7, 14 and
 21. 83. A method of treating cancer ina patient comprising intravenously or orally administering to saidpatient a therapeutically effective amount of a compound represented byformula I:

wherein: Q is selected from the group consisting of

M is selected from the group consisting of oxygen, sulfur, NR⁸, andCR⁹R¹⁰; each R¹, R², R³, R⁴, R⁵, R⁷, R¹¹, R¹², R¹³, R¹⁴ and R¹⁵ is,independently, selected from the group consisting of hydrogen, alkyl,substituted alkyl, aryl, substituted aryl and heterocyclo, and whereinR¹ and R² are alkyl, they can be joined to form a cycloalkyl; R⁶ isselected from the group consisting of hydrogen, alkyl, substitutedalkyl, aryl, substituted aryl, cycloalkyl, heterocyclo and substitutedheterocyclo; R⁸ is selected from the group consisting of hydrogen,alkyl, substituted alkyl, R¹¹C═O, R¹²OC═O and R¹³SO²; and each R⁹ andR¹⁰ is, independently, selected from the group consisting of hydrogen,halogen, alkyl, substituted alkyl, aryl, heterocyclo, hydroxy, R¹⁴C═O,and R¹⁵OC═O; and any salts, solvates, or hydrates thereof.
 84. Themethod of claim 83, wherein the compound of formula I is administeredorally in a dose of about 0.05 mg/kg to 200 mg/kg.
 85. The method ofclaim 83, wherein the compound of formula I is administeredintravenously at a dose of about 1 mg/m² to 65 mg/m².
 86. The method ofclaim 84, wherein the compound is administered daily or weekly.
 87. Themethod of claim 85, wherein the compound is administered daily orweekly.
 88. The method of claim 86 wherein the compound is administereddaily for 3 days with a period of 1 week to 3 weeks between cycles wherethere is no administration of the compound.
 89. The method of claim 87wherein the compound is administered daily for 3 days with a period of 1week to 3 weeks between cycles where there is no administration of thecompound.
 90. The method of claim 86 wherein the compound isadministered daily for 5 days with a period of 1 week to 3 weeks betweencycles where there is no administration of the compound.
 91. The methodof claim 87 wherein the compound is administered daily for 5 days with aperiod of 1 week to 3 weeks between cycles where there is noadministration of the compound.